Sections were then incubated overnight at 4°C with guinea pig anti-insulin (Biogenex ready to use polyclonal antibodies) and mouse antiglucagon (monoclonal clone K79bB10 diluted at 1 : 1000, Sigma) primary antibodies and for 1 hr at room temperature with goat anti-guinea pig Alexa Fluor 488 (Invitrogen) and goat anti-mouse Alexa Fluor 568 (Invitrogen) secondary antibodies diluted in wash buffer at 1 : 300.
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